期刊
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
卷 20, 期 4, 页码 -出版社
MDPI
DOI: 10.3390/ijms20040860
关键词
cell apoptosis; Bcl-2; venetoclax; BH3 mimetics; bioinformatics; in silico; biophysical methods; protein-ligand interactions; high throughput virtual screening
资金
- Applied Molecular Biosciences Unit-UCIBIO - national funds from FCT/MCTES [UID/Multi/04378/2019]
- Fundacao para a Ciencia e a Tecnologia (FCT) Portugal [SFRH/BPD/97719/2013]
Bcl-2 protein is involved in cell apoptosis and is considered an interesting target for anti-cancer therapy. The present study aims to understand the stability and conformational changes of Bcl-2 upon interaction with the inhibitor venetoclax, and to explore other drug-target regions. We combined biophysical and in silico approaches to understand the mechanism of ligand binding to Bcl-2. Thermal shift assay (TSA) and urea electrophoresis showed a significant increase in protein stability upon venetoclax incubation, which is corroborated by molecular docking and molecular dynamics simulations. An 18 degrees C shift in Bcl-2 melting temperature was observed in the TSA, corresponding to a binding affinity multiple times higher than that of any other reported Bcl-2 inhibitor. This protein-ligand interaction does not implicate alternations in protein conformation, as suggested by SAXS. Additionally, bioinformatics approaches were used to identify deleterious non-synonymous single nucleotide polymorphisms (nsSNPs) of Bcl-2 and their impact on venetoclax binding, suggesting that venetoclax interaction is generally favored against these deleterious nsSNPs. Apart from the BH3 binding groove of Bcl-2, the flexible loop domain (FLD) also plays an important role in regulating the apoptotic process. High-throughput virtual screening (HTVS) identified 5 putative FLD inhibitors from the Zinc database, showing nanomolar affinity toward the FLD of Bcl-2.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据